Isolation and Chemical Characterization of Regularly Arrayed Macromolecular Structure in the Cell Wall of Bacillus aneurinolyticus (Kimura et Aoyama)

The tetragonally arrayed macromolecular structure in the cell wall of Bacillus aneurinolyticus (Kimura et Aoyama) S232 was isolated and characterized. The cell wall of B. aneurinolyticus KA was prepared by sonication, differential centrifugation, nuclease treatment, Triton X-100 treatment and washed with water. The regularly arrayed particles were extracted from the purified cell wall preparation with 2M guanidine hydrochloride or 6M urea. SDS-PAGE revealed that the particle of the regular array was a single glycopolypeptide of a molecular weight of 129, 000. The polypeptide contained all amino acids, that were classified into hydrophobic, 39.2% (mole %); neutral, 26.5%; acidic, 21.1%; and basic amino acids, 13.2%. These data indicated the acidic nature of the polypeptide. Its UV absorption spectra in either 6M guanidine hydrochloride or distilled water showed a typical curve of polypeptide with absorption maxima at 277nm and 279nm and an absorption minimum at 250nm in water. The regularly arrayed particles in B. aneurinolyticus KA were compared in the properties with those of other bacterial strains.